WO2023285999A1

WO2023285999A1 - A polyherbal formulation for weight management and a process for its preparation

Info

Publication number: WO2023285999A1
Application number: PCT/IB2022/056482
Authority: WO
Inventors: Sriram Padmanabhan; Vinod Ramachandra Jadhav
Original assignee: Sava Healthcare Ltd
Priority date: 2021-07-15
Filing date: 2022-07-14
Publication date: 2023-01-19

Abstract

The present disclosure relates to a polyherbal formulation. Further, the present disclosure relates to a process of preparing the formulation. The formulation is used for management of 5 obesity and diabetes in the subjects. The formulation is safe and effective.

Description

A POLYHERBAL FORMULATION FOR WEIGHT MANAGEMENT AND A PROCESS FOR ITS PREPARATION

FIELD

The present disclosure relates to a polyherbal formulation and a process for its preparation. Particularly, the present disclosure relates to a polyherbal formulation for weight management.

BACKGROUND

The background information herein below relates to the present disclosure but is not necessarily prior art.

Obesity is a condition in which a person has excess body fat. Obesity is usually defined by using a ratio of height to weight called body mass index (BMI), which often correlates with the person’s level of body fat. A BMI of 30.0 Kg/m or higher is considered as obese in humans. In the current scenario, obesity is very common amongst people due to lack of exercise and disturbed lifestyle which further can increase a person’s risk of diseases and health problems, including high blood pressure, diabetes and heart disease. It is a complex problem and major public health concerns.

Currently, there are various drugs available to decrease the body fat or to control the lipid profile of the subjects. The anti-obesity medications that have been approved by the FDA for chronic weight management are orlistat, phentermine/topiramate, fenfluramine, dexfenfluramine, naltrexone/bupropion, liraglutide and the like. However, they are costly and have life-threatening adverse effects such as cardiac valvulopathy, cancer, stroke, suicidal ideation and behavior and gastrointestinal disturbances like faecal incontinence, diarrhea, constipation, bloating, dyspepsia, dry mouth, poor patient compliance. Due to these serious side effects and complications, the anti-obesity medications are not safe and hence, many of the drugs have been withdrawn from the market.

Further, conventionally the natural plant products are widely used in healthcare or as dietary supplements. The natural plant products contain the important phyto molecules such as flavonoids, terpenoids, saponins, phenols, alkaloids and the like. The phyto molecules would act through their inhibitory activities for pancreatic lipases, adipocyte differentiation or by increasing thermogenesis and anorexia. Many plant-derived molecules have been found to possess anti-obesity effects with advantages over chemical treatments, however, conventional plant-based natural products as anti-obesity therapeutics are largely unexplored and showed comparatively low efficacy.

Therefore, there is felt a need for a polyherbal formulation that mitigates the drawbacks mentioned hereinabove.

OBJECTS

Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows.

An object of the present disclosure is to ameliorate one or more problems of the prior art or to at least provide a useful alternative.

An object of the present disclosure is to provide a polyherbal formulation that is safe.

Another object of the present disclosure is to provide a polyherbal formulation that can be administered orally.

Still another object of the present disclosure is to provide a polyherbal formulation in the form film-coated tablet.

Yet another object of the present disclosure is to provide a polyherbal formulation that is effective in obesity.

Still another object of the present disclosure is to provide a polyherbal formulation which reduces body fat and enhance the lipid profile in obese subjects.

Yet another object of the present disclosure is to provide a process for the preparation of a polyherbal formulation.

Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure. SUMMARY

The present disclosure relates to a polyherbal formulation. The formulation comprises: (a) an extract of Coffee arabica in an amount in the range of 15 wt.% to 20 wt.% with respect to the total weight of the formulation, wherein the extract of Coffee arabica has at least 13% of chlorogenic acid, (b) an extract of Garcinia cambogia in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation, wherein the extract of Garcinia cambogia has at least 50% of hydroxycitric acid, (c) an extract of Terminalia arjuna in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation, wherein the extract of Terminalia arjuna has at least 50% of tannins, (d) an extract of Commiphora mukul in an amount in the range of 2 wt.% to 10 wt.% with respect to the total weight of the formulation, wherein the extract of Commiphora mukul has at least 0.5% of guggulsterone E and Z, (e) an extract of Cyperus rotundus in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation, wherein the extract of Cyperus rotundus has at least 5% of polyphenols, (f) an extract of Zingiber officinale in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation, wherein the extract of Zingiber officinale has at least 0.4 % of polyphenols, (g) an extract of Piper longum in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation, wherein the extract of Piper longum has at least 1% of piperine, (h) an extract of Piper nigrum in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation, wherein the extract of Piper nigrum has at least 2% of piperine, and a pharmaceutically acceptable excipient.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWING

The present disclosure will now be described with the help of the accompanying drawing, in which:

Figure 1 illustrates a UV-Visible chromatogram of mixed standard solution showing peaks of piceatannol and piperine at a wavelength of 340nm;

Figure 2 illustrates a UV-Visible chromatogram of mixed standard solution showing peaks of chlorogenic acid, guggulsterone E and Z at a wavelength of 240nm;

Figure 3 illustrates a UV-Visible chromatogram of sample solution showing peaks of chlorogenic acid, guggulsterone E and Z at a wavelength of 240nm; Figure 4 illustrates a UV-Visible chromatogram of sample solution showing peak of piperine at a wavelength of 340nm;

Figure 5 illustrates a UV-Visible chromatogram of standard solution showing peak of hydroxy citric acid at a wavelength of 215nm;

Figure 6 illustrates a UV-Visible chromatogram of sample solution showing peak of hydroxy citric acid at a wavelength of 215nm;

Figure 7A illustrates a comparison of vital parameters between two treatment groups at different visits as per treatment analysis in accordance with an embodiment of the present disclosure;

Figure 7B illustrates a comparison of vital parameters between two treatment groups at different visits as per protocol analysis in accordance with an embodiment of the present disclosure;

Figure 8 illustrates a change of mean vital parameters across visits in two treatment groups as per protocol analysis;

Figure 9A illustrates a comparison of anthropometric parameters between two groups according to visits as per treatment analysis in accordance with an embodiment of the present disclosure;

Figure 9B illustrates a comparison of anthropometric parameters between two groups according to visits as per protocol analysis in accordance with an embodiment of the present disclosure;

Figure 10 illustrates a comparison of anthropometric parameters across visits in two groups as per protocol analysis in accordance with an embodiment of the present disclosure;

Figure 11 illustrates a comparison of lipid parameters between two treatment groups according to visits in accordance with an embodiment of the present disclosure; and

Figure 12 illustrates a change in mean values of lipid parameters according to visits in two treatment groups in accordance with an embodiment of the present disclosure. DETAILED DESCRIPTION

Embodiments, of the present disclosure, will now be described with reference to the accompanying drawing.

Embodiments are provided so as to thoroughly and fully convey the scope of the present disclosure to the person skilled in the art. Numerous details are set forth, relating to specific components, and methods, to provide a complete understanding of embodiments of the present disclosure. It will be apparent to the person skilled in the art that the details provided in the embodiments should not be construed to limit the scope of the present disclosure. In some embodiments, well-known processes, well-known apparatus structures, and well-known techniques are not described in detail.

The terminology used, in the present disclosure, is only for the purpose of explaining a particular embodiment and such terminology shall not be considered to limit the scope of the present disclosure. As used in the present disclosure, the forms “a,” “an,” and “the” may be intended to include the plural forms as well, unless the context clearly suggests otherwise. The terms "comprises," "comprising," “including,” and “having,” are open ended transitional phrases and therefore specify the presence of stated features, integers, steps, operations, elements, modules, units and/or components, but do not forbid the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. The particular order of steps disclosed in the method and process of the present disclosure is not to be construed as necessarily requiring their performance as described or illustrated. It is also to be understood that additional or alternative steps may be employed.

The most common cause of serious diseases including cancer, neurodegeneration, cardiovascular diseases, diabetes, and kidney diseases is obesity, which in turn results in cell damages and serious alterations in the body and finally lead to serious diseases. Hence, there is an unmet need for a novel formulation that reduces obesity by lowering down the body fat and by maintaining the lipid profile of the subjects. A conventional approach such as chemical treatments via drugs and traditional knowledge does provide options for anti obesity formulations, but they are not efficacious due to their serious adverse effects and poor compliance. There is a definite need for a polyherbal formulation that effectively reduces body fat without having side effects.

The formulation of the present disclosure fills this exact technical gap in the marketplace. The present disclosure provides a polyherbal formulation and a process for its preparation.

In an aspect, the present disclosure provides a polyherbal formulation. The polyherbal formulation comprises (a) an extract of Coffee arabica in an amount in the range of 15 wt.% to 20 wt.% with respect to the total weight of the formulation, wherein the extract of Coffee arabica has at least 13% of chlorogenic acid, (b) an extract of Garcinia cambogia in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation, wherein the extract of Garcinia cambogia has at least 50% of hydroxycitric acid, (c) an extract of Terminalia arjuna in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation, wherein the extract of Terminalia arjuna has at least 50% of tannins, (d) an extract of Commiphora mukul in an amount in the range of 2 wt.% to 10 wt.% with respect to the total weight of the formulation, wherein the extract of Commiphora mukul has at least 0.5% of guggulsterone E and Z, (e) an extract of Cyperus rotundus in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation, wherein the extract of Cyperus rotundus has at least 5 % of polyphenols, (f) an extract of Zingiber officinale in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation, wherein the extract of Zingiber officinale has at least 0.4 % of polyphenols, (g) an extract of Piper longum in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation, wherein the extract of Piper longum has at least 1% of piperine, (h) an extract of Piper nigrum in an amount ranging from 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation, wherein the extract of Piper nigrum has at least 2% of piperine, and a pharmaceutically acceptable excipient.

In an embodiment, the plant extract is derived from barks, roots, tubers, stolons, rhizomes, leaves, seeds, fruits, stems and flowers.

In an embodiment, the extracts of Coffee arabica, Garcinia cambogia, Terminalia arjuna, Commiphora mukul, Cyperus rotundus, Zingiber officinale, Piper longum, Piper nigrum is taken in the form of a powder obtained by direct micronization of the plant material. Alternatively, the extracts may be in the form of a solid or a semi-solid or liquid. Typically, the extracts are selected from the group consisting of alcoholic, hydroalcoholic, aqueous, ether, ester, ethyl acetate, acetone and hexane extract. Typically, the extracts are prepared by using techniques selected from percolation, decoction, maceration, Soxhlet extraction and supercritical fluid extraction. Coffee arabica is also known as the Arabian coffee, "coffee shrub of Arabia", "mountain coffee" or "arabica coffee", is a species of Coffee. Arabica coffee originates from Ethiopia. Extracts of seeds have high anti-obesity functions which exert lipolytic and thermogenic actions.

In accordance with the embodiments of the present disclosure, the amount of the extract of Coffee arabica is in the range of 15 wt. % to 20 wt. % of the total weight of the formulation wherein the extract of Coffee arabica has at least 13% of chlorogenic acid. In an exemplary embodiment, the amount of extract of Coffee arabica is 18.22 wt. % wherein the extract has 13% of chlorogenic acid.

Garcinia cambogia, a tropical fruit also known as the Malabar tamarind. The active ingredient hydroxy citric acid (HCA) in the fruit’s rind has fat-burning property and also put back to have low appetite. It appears that hydroxycitric acid (HCA) blocks an enzyme called citrate lyase, which the body uses to make fat.

In accordance with the embodiments of the present disclosure, the amount of the extract of Garcinia cambogia is in the range of 5 wt. % to 15 wt. % of the total weight of the formulation wherein the extract of Garcinia cambogia has at least 51% of hydroxycitric acid. In an exemplary embodiment, the amount of extract of Garcinia cambogia is 10.41 wt. % wherein the extract has 50% of hydroxycitric acid.

Arjuna ( Terminalia arjuna ) is a tree of the genus Term in alia. It is commonly known as arjuna or arjun tree in English. The arjuna is seen across the Indian Subcontinent, and usually found growing on river banks or near dry river beds in Uttar Pradesh, Bihar, Maharashtra, Madhya Pradesh, West Bengal, Odisha and south and central India, along with Sri Lanka and Bangladesh. It has also been planted in Malaysia, Indonesia and Kenya. It contains ellagic acid, arjunolic acid arjunine, triterpene glycosides like arjunetosides I, II, III & IV. The bark is rich in saponins and natural anti-oxidants which have been reported to reduce cholesterol levels. Terminalia Arjuna increases cardiac muscle functions and pumping action of the heart.

In accordance with the embodiments of the present disclosure, the amount of the extract of Terminalia arjuna is in the range of 5 wt. % to 15 wt. % of the total weight of the formulation wherein the extract of Terminalia arjuna has at least 50 % of tannins. In an exemplary embodiment, the amount of the extract of Terminalia arjuna is 7.81 wt.% wherein the extract of Terminalia arjuna has 50 % of tannins.

Guggul (Commiphora mukul or wightii) or Balsamodendron mukul with common names the Indian bdellium- tree, gugal, guggul, guggul, or Mukul myrrh tree, is a flowering plant in the family Burseraceae, which produces a fragrant resin called gugal, guggul or gugul, that is used in incense and vedic medicine (or Ayurveda). The guggul plant may be found from northern Africa to central Asia, but is most common in northern India. It prefers arid and semi-arid climates and is tolerant of poor soil. Balsamodendron mukul enhances the body’s metabolic activity by increasing the fat burning activity and by heat production. Extracts containing ketonic steroids show significantly lower serum low-density lipoprotein and very low-density lipoprotein and triglycerides.

In accordance with the embodiments of the present disclosure, the amount of the extract of Commiphora mukul is in the range of 2 wt. % to 10 wt. % of the total weight of the formulation wherein, the extract of Commiphora mukul has at least 0.5% of guggulsterone E and Z. In an exemplary embodiment, the amount of the extract of Commiphora mukul is 5.85 wt. % wherein the extract of Commiphora mukul has 0.5% of guggulsterone E and Z.

Nagarmoth ( Cyperus rotundus ) or coco-grass, java grass, nutgrass, purple nutsedge or purple nutsedge, red nut sedge, Khmer kravanh chruk is a species of sedge native to Africa, southern and central Europe (north to France and Austria), and southern Asia. Cyperus rotundus belonging to the family Cyperaceae is a well-known plant in the Indian traditional medicine prescribed for exerting anti-obesity. Cyperus rotundus regulates serum lipid profile, reduces oxidative stress and decreases adipose tissue mass and body weight gain.

In accordance with the embodiments of the present disclosure, the amount of the extract of Cyperus rotundus is in the range of 5 wt. % to 15 wt. % of the total weight of the formulation wherein the extract of Cyperus rotundus has at least 5% of polyphenols. In an exemplary embodiment, the amount of the extract of Cyperus rotundus is 7.16 wt. % wherein the extract of Cyperus rotundus has 5% of polyphenols.

Ginger ( Zingiber officinale) or shunthi belongs to the family Zingiberaceae, is a flowering plant whose rhizome, ginger root or ginger, and is widely used as a spice and folk medicine. It is an herbaceous perennial which grows annual pseudostems (false stems made of the rolled bases of leaves) about one-meter tall bearing narrow leaf blades. Ginger is originated in Island Southeast Asia and was likely domesticated first by the Austronesian peoples. The plant has several chemicals that is responsible for its medicinal properties, such as anti- arthritis, anti-inflammatory, anti-diabetic, anti-bacterial, anti-fungal, and anti-cancer, and the like. Zingiber officinale enhances anthropometric measurements and hence reduces obesity in the subjects. It reduces obesity through various potential mechanisms including increasing thermogenesis, increasing lipolysis, suppression of lipogenesis, inhibition of intestinal fat absorption, and controlling appetite.

In accordance with the embodiments of the present disclosure, the amount of the extract of Zingiber officinale is in the range of 0.5 wt. % to 5 wt. % of the total weight of the formulation wherein the extract of Zingiber officinale has at least 0.4% of polyphenols. In an exemplary embodiment, the amount of the extract of Zingiber officinale is 1.3 wt. % of the total weight of the formulation wherein the extract of Zingiber officinale has 0.4% of polyphenols.

Pipalimul ( Piper longum) sometimes called the Indian long pepper or pipli, is a flowering vine in the family Piperaceae, cultivated for its fruit, which is usually dried and used as a spice and seasoning. Piper longum contains an alkaloid piperine (5%), essential oils like piperlongumine, piperlonguminine. It is a well-known antioxidant, bioavailability enhancer and thermogenic agent and proven a significant decrease in cholesterol and increase in HDL levels in studies.

Piper longum is anushna (neutral) and has a post-digestive effect which is less irritatable to the GI tract as compared to Piper nigrum.

In accordance with the embodiments of the present disclosure, the amount of the extract of Piper longum is in the range of 0.5 wt. % to 5 wt. % of the total weight of the formulation wherein the extract of Piper longum has at least 1% of piperine. In an exemplary embodiment, the amount of the extract of Piper longum is 1.3 wt. % of the total weight of the formulation wherein the extract of Piper longum has 1 % of piperine.

Mari ( Piper nigrum) is a flowering vine in the family Piperaceae, cultivated for its fruit, known as a peppercorn. Piper nigrum has anti-obesity and anti-hyperglycemic activities and shown a significant decrease in cholesterol and increase in HDL levels in studies. Piper nigrum is known for generating heat (ushna) and causes pitta. In accordance with the embodiments of the present disclosure, the amount of the extract of Piper nigrum is in the range of 0.5 wt. % to 5 wt. % of the total weight of the formulation wherein the extract of piper nigrum has at least 2% of piperine. In an exemplary embodiment, the amount of the extract of Piper nigrum is 1.3 wt. % of the total weight of the formulation wherein the extract of piper nigrum has 2% of piperine

In accordance with the embodiments of the present disclosure, at least one pharmaceutically acceptable excipient is selected from the group consisting of a disintegrant, a binder, a preservative, a fluid medium , a glidant, a film-forming agent, and a coloring agent.

In accordance with the embodiments of the present disclosure, the disintegrants is at least one selected from the group consisting of polyvinylpyrrolidone (crospovidone), sodium starch glycolate, sodium croscarmellose, starch, and colloidal anhydrous silica. In an exemplary embodiment, the disintegrant is a combination of polyvinylpyrrolidone (crospovidone) and sodium starch glycolate.

In accordance with the embodiments of the present disclosure, the amount of the disintegrant is in the range of 15 to 20% with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the disintegrant is 18.22 wt.% with respect to the total weight of the formulation.

In accordance with the embodiments of the present disclosure, the binder is at least one selected from microcrystalline cellulose, povidone, hydroxymethyl cellulose, ethyl cellulose, hydroxypropyl cellulose, starch, gum acacia, alginate, and carboxymethyl cellulose. In an exemplary embodiment, the binder is microcrystalline cellulose.

In accordance with the embodiments of the present disclosure, the amount of the binder is in the range of 20 to 30% with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the binder is 24 wt.% with respect to the total weight of the formulation.

In accordance with the embodiments of the present disclosure, the preservative is at least one selected from the group consisting of methyl hydroxybenzoate (methylparaben), propyl hydroxybenzoate (propylparaben), ethyl paraben, and sodium benzoate. In an exemplary embodiment, the preservative is a combination of methylparaben and propylparaben. In accordance with the embodiments of the present disclosure, the amount of the preservative ranging from 0.01 to 0.05% with respect to the total weight of the excipients. In an exemplary embodiment, the amount of the preservative is 0.03 wt.% with respect to the total weight of the excipients.

In accordance with the embodiments of the present disclosure, the fluid medium is at least one selected from the group consisting of isopropanol (isopropyl alcohol), ethanol, methanol, hexane, polyethylene glycol 4000, and dichloromethane (methylene chloride). In an exemplary embodiment, the fluid medium is isopropyl alcohol. In another exemplary embodiment, the fluid medium is polyethylene glycol 4000. In still another exemplary embodiment, the fluid medium is dichloromethane.

In accordance with the embodiments of the present disclosure, the glidant is at least one selected from silicon dioxide (silica), magnesium stearate, purified talc, calcium palmitate, ascorbyl palmitate, and starch. In an exemplary embodiment, the glidant is a combination of silica, magnesium stearate, and talc.

In accordance with the embodiments of the present disclosure, the amount of the glidant is in the range of 1 to 5% with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the glidant is 2.3 wt.% with respect to the total weight of the formulation.

In accordance with the embodiments of the present disclosure, the film-forming agent is at least one selected from hydroxypropyl methylcellulose, ethyl cellulose, hydroxypropyl cellulose, povidone, and polyvinyl alcohol. In an exemplary embodiment, the film-forming agent is hydroxypropyl methylcellulose.

In accordance with the embodiments of the present disclosure, the amount of the film- forming agent is in the range of 1 to 5% with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the film-forming agent is 1.77 wt.% with respect to the total weight of the formulation.

In accordance with the embodiments of the present disclosure, the coloring agent is at least one selected from the group consisting of titanium dioxide, iron oxide red, iron oxide black, tartrazine yellow, indigotine, sunset yellow, and brilliant blue. In an exemplary embodiment, the coloring agent is a combination of titanium dioxide, iron oxide red, and iron oxide black. In accordance with the embodiments of the present disclosure, the amount of the coloring agent is in the range of 0.1 to 0.5% with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the coloring agent is 0.26wt.% with respect to the total weight of the formulation.

In an exemplary embodiment, the formulation comprises 18.22 wt.% extract of Coffee arabica, 10.41 wt.% extract of Garcinia cambogia, 7.8 wt.% extract of Terminalia arjuna 5.85 wt.% extract of Commiphora mukul, 7.16 wt.% extract of Cyperus rotundas, 1.3 wt.% extract of Zingiber officinale, 1.3 wt.% extract of Piper longum, 1.3 wt.% extract of Piper nigrum, 23.95 wt.% of microcrystalline cellulose (Avicel PH 101), 9.7wt.% of crospovidone, 8.46 wt.% of sodium starch glycolate, 61.5 wt.% of isopropyl alcohol, 0.02 wt.% of methyl paraben, 0.005 wt.% of propyl paraben, 1.17 wt.% silica (syloid 244 FP), 0.9 wt.% of magnesium stearate, 1.77 wt.% of hydroxypropyl methyl cellulose, 0.23 wt.% of polyethylene glycol, 0.04 wt.% of titanium dioxide, 0.04 wt.% of talc, 0.1 wt.% of iron oxide red, 0.15 wt.% iron oxide black, and 22.4 wt.% of methylene chloride.

In an embodiment of the present disclosure, the formulation is in the form of a tablet selected from uncoated tablet, film -coated tablet, sugar-coated tablet, and enteric-coated tablet.

In an exemplary embodiment, the formulation is a film-coated tablet.

In another aspect, there is provided with a process for the preparation of the polyherbal formulation in the form of a tablet. The process is described in details as:

In a first step, sifting individually powder extracts of Coffee arabica, Garcinia cambogia, Terminalia arjuna, Commiphora mukul, Cyperus rotundas, Zingiber officinale, Piper longum and Piper nigrum through a sieve no. 20 to obtain a sieved extract. Similarly, sifting individually a binder, a disintegrant through a sieve no. 40 and glidant through a sieve no. 60 to obtain a sieved binder, glidant, and disintegrant. The sieved extracts of Coffee arabica, Garcinia cambogia, Terminalia arjuna, Commiphora mukul, Cyperus rotundas, Zingiber officinale, Piper longum and Piper nigrum are then mixed with a sieved binder, glidant, and disintegrant in the rapid mixer granulator (RMG) under stirring to obtain a first mixture;

In the second step, separately, a predetermined amount of at least one fluid medium and a predetermined amount of at least one preservative are mixed in a vessel to obtain a solution. In an embodiment of the present disclosure, the fluid medium is at least one selected from the group consisting of isopropanol (isopropyl alcohol), ethanol, methanol, hexane, polyethylene glycol 4000, and dichloromethane (methylene chloride). In an exemplary embodiment, the fluid medium is isopropyl alcohol.

In an embodiment of the present disclosure, the preservative is at least one selected from the group consisting of methyl hydroxybenzoate (methylparaben), propyl hydroxybenzoate (propylparaben), ethyl paraben, and sodium benzoate. In an exemplary embodiment, the preservative is a combination of methylparaben and propylparaben.

In an embodiment of the present disclosure, the predetermined amount of extract of Coffee arabica is in the range of 15 wt.% to 20 wt.%, the predetermined amount of extract of Garcinia cambogia is in the range of 5 wt.% to 15 wt.%, the predetermined amount of extract of Terminalia arjuna is in the range of 5 wt.% to 15 wt.%, the predetermined amount of extract of Commiphora mukul is in the range of 2 wt.% to 10 wt.%, the predetermined amount of extract of Cyperus rotundas is in the range of 5 wt.% to 15 wt.%, the predetermined amount of extract of Zingiber officinale is in the range of 0.5 wt.% to 5 wt.%, the predetermined amount of extract of Piper longum is in the range of 0.5 wt.% to 5 wt.%, the predetermined amount of extract of Piper nigrum is in the range of 0.5 wt.% to 5 wt.%, the predetermined amount of binder is in the range of 20 to 25% with respect to the weight of the formulation, the predetermined amount of glidant is in the range of 1 to 5% with respect to the weight of the formulation, the predetermined amount of disintegrant is in the range of 15 to 20% with respect to the weight of the formulation, and the predetermined amount of preservative is in the range of 0.01 to 0.05% with respect to the weight of the formulation.

In the third step, the solution is added into the first mixture in the mixer under stirring for a first predetermined time period to obtain a second mixture.

In an embodiment of the present disclosure, the first predetermined time period is in the range of 3 to 10 min. In an exemplary embodiment, the first predetermined time period is 5 min.

In the fourth step, the second mixture is kneaded under stirring for a second predetermined time period followed by granulation to obtain the granules. In an embodiment of the present disclosure, the second predetermined time period is in the range of 4 to 8 min. In an exemplary embodiment, the second predetermined time period is 6 min.

In the fifth step, the so obtained granules are dried in the dryer (fluid bed dryer) till the loss of drying is achieved at a predetermined temperature to obtain dried granules.

In an embodiment of the present disclosure, the predetermined temperature is in the range of 40 to 50°C. In an exemplary embodiment, the predetermined temperature is 45 °C.

In the sixth step, the dried granules are sieved through a sieve no. 20 and further the retained granules are sieved through a 2 mm perforated stainless-steel screen to obtain milled granules followed by sieving the milled granules through sieve no 16 to obtain uniform milled granules.

In the seventh step, the milled granules are mixed with a predetermined amount of disintegrant and a predetermined amount of binder for a third predetermined time period to obtain a blend.

In accordance with the embodiments of the present disclosure, the disintegrant is at least one selected from the group consisting of polyvinylpyrrolidone (crospovidone), sodium starch glycolate, sodium croscarmellose, starch, and colloidal anhydrous silica. In an exemplary embodiment, the disintegrant is a combination of polyvinylpyrrolidone (crospovidone) and sodium starch glycolate.

In accordance with the embodiments of the present disclosure, the binder is at least one selected from the group consisting of microcrystalline cellulose, povidone, hydroxymethyl cellulose, ethyl cellulose, hydroxypropyl cellulose, starch, gum acacia, alginate, and carboxymethyl cellulose. In an exemplary embodiment, the binder is microcrystalline cellulose.

In accordance with the embodiments of the present disclosure, the amount of the binder ranging from 20 to 25% with respect to the total weight of the excipients. In an exemplary embodiment, the amount of the binder is 24 wt.% with respect to the total weight of the excipients.

In an embodiment of the present disclosure, the third predetermined time period is in the range of 8 to 12 min. In an exemplary embodiment, the third predetermined time period is 10 min.

In the eighth step, the blend further mixed with a predetermined amount of at least one glidant for a fourth predetermined time period to obtain lubricated granules.

In accordance with the embodiments of the present disclosure, the glidant is at least one selected from silicon dioxide (silica), magnesium stearate, talc, calcium palmitate, ascorbyl palmitate, and starch. In an exemplary embodiment, the glidant is a combination of silica and magnesium stearate.

In accordance with the embodiments of the present disclosure, the amount of the glidant ranging from 1 to 5 % with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the glidant is 2.3 wt.% with respect to the total weight of the formulation.

In an embodiment of the present disclosure, the fourth predetermined time period is in the range of 2 to 10 min. In an exemplary embodiment, the fourth predetermined time period is 5 min.

In the ninth step, the lubricated granules are punched to obtain the uncoated tablet.

The so obtained uncoated tablet is coated with a predetermined amount of a layer of coating solution, followed by adding a predetermined amount of at least one glidant and a coloring agent to obtain a film-coated tablet.

In accordance with the embodiments of the present disclosure, the coating solution is comprises hydroxypropyl methylcellulose, polyethylene glycol, titanium dioxide, talc, colors, and solvents.

In accordance with the embodiments of the present disclosure, the coloring agent is at least one selected from titanium dioxide, iron oxide red, iron oxide black, tartrazine yellow, indigotine, sunset yellow, and brilliant blue. In an exemplary embodiment, the coloring agent is a combination of titanium dioxide, iron oxide red, and iron oxide black. In accordance with the embodiments of the present disclosure, the amount of the coloring agent is in the range of 0.1 to 0.5% with respect to the total weight of the formulation. In an exemplary embodiment, the amount of the coloring agent is 0.26 wt.% with respect to the total weight of the formulation.

The polyherbal composition of the present disclosure is effective in controlling obesity and diabetics in the human. Moreover, the polyherbal composition of the present disclosure is safe.

The foregoing description of the embodiments has been provided for purposes of illustration and not intended to limit the scope of the present disclosure. Individual components of a particular embodiment are generally not limited to that particular embodiment, but, are interchangeable. Such variations are not to be regarded as a departure from the present disclosure, and all such modifications are considered to be within the scope of the present disclosure. The present disclosure is further described in light of the following experiments which are set forth for illustration purpose only and not to be construed for limiting the scope of the disclosure. The following experiments can be tested to scale up to industrial/commercial scale and the results obtained can be extrapolated to the industrial scale.

EXPERIMENTAL DETAILS: EXPERIMENT 1: Preparation of the polyherbal formulation in the form of a film- coated tablet:

The polyherbal formulation in the form of a tablet was prepared by blending the ingredients and appropriate quantities as given in Table- 1

Table 1

EXPERIMENT 2: Assay determination of active constituents by using Chromatogram

Active constituents such as chlorogenic acid, piceatannol, piperine, guggulsterone E, guggulsterone Z and hydroxycitric acid were assayed by using chromatogram and peak is measured for each constituent.

Chromatographic conditions:

Buffer solution Preparation: 1.36 gm of Potassium dihydrogen phosphate was dissolved in 900mL of water and pH was adjusted to 2.5 with 30% of phosphoric acid, final volume was made up to 1000 mL with water. The buffer solution was mixed, degassed and filtered through a 0.45μ filter.

Mobile Phase A: Buffer solution.

Mobile Phase B: Methanol.

Column : Inertsil ODS, 3V, 250 x 4.6 mm and 5μm. Column Temperature: 30°C

Wave length: hydroxycitric acid- 215nm, Chlorogenic acid, Guggulsterone E and Guggulsterone Z - 240nm, and Piceatannol, Piperine-340nm.

Flow rate: 1.0 ml, /min

Injection Volume: 20 μL Run time: 60 minutes

Retention Time: Hydroxycitric acid about 4.5 min, Chlorogenic acid at about 25 min., Piceatannol at about 32 min, Piperine at about 50 min, Guggulsterone E at about 53 min, and Guggulsterone Z at about 54 min.

Elution : Gradient Gradient Program: Table 2

Preparation of individual Standard stocks: Chlorogenic acid Standard stock: (100μg / mL)

2 mg of the chlorogenic acid reference standard (Natural remedies) was transferred to 20 mL volumetric flask. 10 mL of 50% ethanol (in water) was added and sonicated to dissolve the chlorogenic acid. The final volume was made up to the mark with 50% ethanol (in water) and mixed well to obtain standard stock solution of chlorogenic acid. Piceatannol Standard stock: (100μg / mL)

2 mg of the piceatannol reference standard (TCI) was transferred into 20 mL volumetric flask. 10 mL of 50% ethanol (in water) was added and sonicated to dissolve the piceatannol. The final volume was made up to the mark with 50% ethanol (in water) and mixed well to obtain standard stock solution of Piceatannol. Piperine Standard stock: (50μg / mL)

2 mg of Piperine reference standard (Sigma) was transferred into 20 mL volumetric flask. 15 mL of 100% methanol was added and sonicated to dissolve the Piperine. The final volume was made up to the mark with 100% methanol and mixed well. Further, 5mL of the solution was taken from the final volume and diluted with 10ml of methanol (100%) followed by mixing to obtain the piperine standard stock solution. Guggulsterone E Isomer and Guggulsterone Z Isomer Standard stock: (each 10μg / mL)

2 mg of Guggulsterone E reference standard (Natural remedies) and 2 mg of Guggulsterone Z reference standard (Natural remedies) were transferred into 20 mL volumetric flask. 10 mL of methanol (100%) was added and sonicated to dissolve the Guggulsterone E and the Guggulsterone Z. The final volume was made up to the mark with 100% methanol and mixed well. Further, 1mL of the solution was taken from the final volume and diluted with 10ml of methanol (100%) followed by mixing to obtain the Guggulsterone E Isomer and Guggulsterone Z Isomer Standard stock solution.

Mix Standard preparation (Chlorogenic acid 10μg / mL, Piceatannol 10μg / mL, Piperine 5μg / mL Guggulsterone E 1μg / mL, Guggulsterone Z 1μg / mL)

1 mL Standard stocks of each chlorogenic acid, piceatannol, piperine, guggulsterone E and guggulsterone Z Isomer were taken into lOmL volumetric flask and diluted up to the mark with 50% methanol (in water) and mixed well.

Hydroxycitric acid standard solution: (50μg / mL)

25 mg of hydroxycitric acid reference standard /working standard was transferred into 10 mL volumetric flask. 5 mL of 3% orthophosphoric acid (in water) was added and sonicated to dissolve hydroxycitric acid. The final volume was made with 3% orthophosphoric acid (in water) and mixed well. Lurther, diluted lmL of the above solution to 50mL with 3% orthophosphoric acid (in water) mixed well to obtain hydroxycitric acid standard solution.

Sample Preparation- 1 (for Chlorogenic acid, Piceatannol, Piperine, Guggulsterone E, Guggulsterone Z):

At least 20 tablets were taken randomly and an average weight of tablets was calculated and crushed into fine powder. 100 mg powder of tablets was transferred to 100 mL volumetric flask and 60 mL of 50% methanol (in water) was added and sonicated for 30 minutes with intermediate shaking. Made the final volume with 50% methanol (in water) and mixed well. The solution was filtered through a 0.45 m nylon filter and the filtrate was collected by discarding the first 3 mL of the filtrate.

Sample Preparation-2 (for Hydroxy citric acid): At least 20 tablets were taken randomly and an average weight of tablets was calculated and crushed into fine powder. 100 mg powder of tablets was transferred to 100 mL volumetric flask and 60 mL of 3% orthophosphoric acid (in water) was added and sonicated for 30 minutes with intermediate shaking. Made the final volume with 3% orthophosphoric acid (in water) and mixed well. The solution was filtered through 0.45 m nylon filter and the filtrate were collected by discarding the first 3 mL of the filtrate.

System Suitability:

The chromatograph was conducted for standard preparations and the multiple peak responses was recorded as illustrated in figures 1 to 6. Retention time, peak type, and area of each active constituent was recorded which is mentioned below in table 3:

Table 3:

Inference: It is evident from table 3 and figures 1 to 6 that extracts and formulation of the present disclosure shows the presence of the active constitutes at a specific wavelength.

Procedure: Separately injected the diluent, standard preparation and sample preparation into the chromatograph. The response for the analyte peaks was measured and recorded into the chromatograms.

The quantity in mg per tablets and % assay was calculated of each content using the following formula: Calculation:

Chlorogenic

Acid

(mg/tablet)

Piceatannol

(mg/tablet)

Piperine

(mg/tablet)

Guggulsterone E (mg/tablet)

Guggulsterone Z (mg/tablet)

Hydroxycitric

Acid

(mg/tablet)

% Assay

where, At: Average area of analyte peak in Sample Solution As: Average area of analyte peak in Standard Solution Wt. std. : Weight of standard in mg Wt. spl: Weight of sample in mg Avg.Wt: Average weight in mg P: Potency of Standard in % (as such basis) L.C. : Label Claim in mg

EXPERIMENT 3: Evaluation of the efficacy and safety of the polyherbal formulation in obese subjects:

The safety and efficacy of the polyherbal formulation of the present disclosure in obese subjects were evaluated. The clinical trial was conducted as per the protocol design and ICH- GCP guidelines.

A total of 120 obese subjected were considered for this study out of which 60 males and 60 females having age group of 18 to 65 years were selected in two groups with body mass index (BMI) between 25-34.9 Kg/m² for the evaluation of the formulation. One group of the subjects received the formulation of the present disclosure and the other group of subjects received the placebo-controlled drug. The route of administration was oral and the frequency of dose was two tablets twice a day after each meal (three meals in total) for 90 days. The study was designed as a multicenter, double-blind, placebo-controlled, interventional and observational study. Parameters such as anthropometric measurements, leptin level, quality of life questionnaires, and safety parameters like hematology and biochemistry were assessed to evaluate the formulation. The total duration of the study was 8 months. To evaluate the safety and efficacy of the polyherbal formulation of the present disclosure, the primary and the secondary objectives were assessed and adverse events were also reported for the safety.

The primary objective of the study was to assess the change in body weight (kg) over 3 months of the study.

Secondary objectives were assessed by looking at the following: change in weight, change in anthropometric measurements, change in lipid profile, change in serum leptin level, and change in the quality of life.

Safety was assessed by looking at adverse events and tolerability of the polyherbal formulation.

Inclusion criteria: male and/or female volunteers aged between 18 to 65 years. > BMI ( > ) greater than or equal to 25 Kg/m² and less than or equal to (<) 34.9 Kg/m².

> willing to do exercise for 30 minutes daily for at least 5 days a week.

> willing to follow the diet advised by the investigator.

> willing to come for regular follow-up visits.

Exclusion criteria:

> intake of over-the-counter weight loss agents, centrally acting appetite suppressants, or prior surgery for obesity in the previous three months.

> history of pathophysiologic/genetic syndromes associated with obesity (Cushing’s syndrome, Turner’s syndrome, and Parder- Willi Syndrome).

> alcoholics and subjects with substance abuse.

> subject with evidence of malignancy

> subjects having a history of underlying inflammatory arthropathy, septic arthritis, inflammatory joint disease, gout, pseudo gout, Paget’s disease, joint fracture, acromegaly, fibromyalgia, and rheumatoid arthritis.

> subjects having a history of coagulopathies, cardiovascular diseases, and asthma.

> subjects with HbAlc> 7 % and poorly controlled diabetes mellitus, and poorly controlled hypertension.

> TSH > 10 mlU/L and T3 and T4 higher than the normal range.

> pregnant and lactating women.

> subjects with hepatic and renal failure.

> subjects on prolonged (> 6 weeks) medication with corticosteroids, antidepressants, anticholinergics, etc., or any other drugs that may influence the outcome of the study.

> subjects with hypersensitivity to any one ingredient of the drugs.

> subjects who have participated in another trial within the last 3 months.

> any other condition which the principal investigator thinks may jeopardize the study.

Screening of the subjects: The subjects were screened before the treatment to obtain a bias- free data. All the included subjects (120) were gone through the screening visits at a specific interval of time. The investigator had a screening visit 1 (day 10), a baseline visit 2 (day 0), a visit 3 (day 30 ±5), a visit 4 (day 60 ± 5), a visit 5 (day 90 ± 5), and a follow-up visit.

On the screening visit 1 (day 10) the investigator checked inclusion and exclusion criteria of subjects, disease history, present signs, and symptoms, and also recorded demographics (age, gender, height, weight (screening only)) and vital signs (Pulse Rate, Respiratory Rate, Temperature, BP), blood samples, quality of life questionnaire, BMI, anthropometric measurements like waist circumference, hip circumference, waist: hip ratio, body fat percentage, basal metabolic rate (BMR), total body water (TBW), lean body mass (LBM), neck circumference, upper-mid-arm circumference, calf circumference to check the eligibility criteria for all the subjects.

On the baseline visit 2 (day 0) the investigator analyzed the quality of life (QOL) questionnaires BMI, lab reports (CBC, physical examinations, urine, and vital signs), anthropometric measurements, and tablets were prescribed for 30 days along with the exercise and the diet plan till day 30.

On the screening visit 3 (day 30 ±5), the screening visit 4 (day 60 ± 5), the screening visit 5 (day 90 ± 5), the investigator further analyzed the quality of life (QOL) questionnaires BMI, lab reports (CBC, physical examinations, urine, and vital signs), anthropometric measurements and tablets were prescribed for remaining 60 days respectively along with exercise and diet plan till day 90. Further, any adverse event or serious adverse events experienced for the past month from screening to date along with the compliance was also recorded.

In the follow-up visit, the investigator had a telephonic conversation for at least 15 days from the last visit where subjects were called for inquiry about his/her wellbeing on the post drug treatment. Subjects were asked for any adverse events experienced since the last study visit and the same was noted in the telephonic log.

Clinical evaluation of the efficacy of the polyherbal formulation by using a statistical analysis:

All data were reported as per the protocol and the treatment were summarized in the statistical tools to evaluate the efficacy of the polyherbal formulation of the present disclosure in the obese subjects.

All the parameters were measured by using the statistical tools and recorded at the time of screening the subjects which are mentioned below: • Vital parameters: various vital parameters such as systolic BP (mmHg), diastolic BP(mmHg), pulse (beats/min), heart rate ((beats/min), respiratory rate (per min), temperature (°F), weight (kg), height (cm), and BMI (Kg/m² ) of the patients were reported normal.

• Urine pregnancy test: Out of 80 females screened, the pregnancy test was carried out in 63 females, out of which only one was positive, while 62 (98.4%) were negative. There were 2 (2.5%) females in which the test was not performed, 8 (10%) were menopausal cases and 7 (8.7%) were surgically sterile. · General physical parameters such as appearance, ear, nose, throat, head, heart, lungs, abdomen, neurological, and extremities were found normal.

• Patients were screened for their past medical and medication history. There were 7 (5.4%) cases of cardiovascular disease, followed by 5 (3.8%) cases of endocrine disorder. There were 3 (2.3%) cases of genitourinary, while 2 (1.5%) cases of gastrointestinal disorder. Other occurrences were observed in less than 1% of the cases. Further, there were 12 (9.23%) cases with medication history at the time of screening. Only 2 (1.5%) patients were on the Ayurvedic treatment.

• All the lab parameters and anthropometric parameters were also found to be normal at the time of screening the subjects.

Comparison of vital parameters between two treatment arms (formulation- 252A and placebo-252B) at baseline and subsequent visits as per the treatment protocol analysis is mentioned in table 4A:

5 All the randomized patients were considered for the analysis. At baseline, 60 patients were randomized to each treatment arm, while at visit #3, there were 5 dropouts in arm 252 A and 2 in arm 252B. Further, at visit #4, there was one drop out in arm 252 A and 3 in arm 252B. In visit #5, there was no drop out in either of the ar s. The comparison of parameters between two groups was performed by 10 using a t-test for independent samples at each time point.

Table 4A

wherein *P-value calculated by using t-test for independent samples and bold p-value indicate statistical significance.

Inference: It is evident from table 4A and figures 7A that at baseline, none of the 5 parameters showed a statistically significant difference of means between the two groups (p-value > 0.05). Similar was the observation at visit #3 and visit #4. At visit #5, mean diastolic BP in arm 252A (77.19 ± 6.34 mmHg) was significantly higher than that of arm 252B (74.64 ± 5.30 mmHg) as indicated by p-value of 0.025 (p < 0.05). Other parameters were insignificantly different between the two 10 groups.

Comparison of vital parameters between two treatment arms at baseline and subsequent visits as per the protocol is mentioned in table 4B:

Table 4B

Patients with complete information till the last visit were considered for analysis. 15 There were 54 patients with complete details in arm 252A, while 55 patients were in arm 252B.

Inference: It is evident from the descriptive statistics of table 4B and figure 7B that 5 baseline and visit #3 showed differences as compared to table 3A (treatment) analysis due to a change in the number of patients (n). However, the comparison of vital parameters with the per-protocol sample showed a statistically insignificant difference of means between the two groups as indicated by p-values > 0.05. Similar was the observation at visit #3. The results for visits #4 and #5 were similar as above 10 in table 3 A.

Comparison of vital parameters across visits in each study group (n) 15 (formulation-252A and placebo-252B) as per the protocol analysis is mentioned in table 5: Table 5: provides the comparison of vital parameters across times in two treatment arms on a per protocol basis using repeated measure analysis of variance.

wherein *P-value calculated by using repeated measure analysis of variance 5 (ANOVA). Bold p-values indicate the statistical significance.

Inference: It is evident from the descriptive statistics of table 5 and figure 8 that in arm 252A, systolic, diastolic BP, heart rate, and temperature did not show statistically significant differences across times (p-value > 0.05). The mean pulse rate showed a significant difference across times (p-value: 0.003). The mean at visit 10 #4 (81.89 ± 6.93 beats/min) was significantly higher at visit #4 compared to baseline and visit #3. The mean was marginally reduced to 81.13 ± 6.83 beats/min at visit #5. The respiratory rate also showed a statistically significant difference across visits (p- value: 0.001). The mean respiratory rate at visit #3 (18.86 ± 1.92 /min) was significantly higher as compared to baseline. However, the mean was reduced at visit 15 #4 and #5 marginally as compared to visit #3.

In arm 252B, systolic BP varied insignificantly across times; however, diastolic BP showed a statistically significant change of means across visits (p-value: 0.003). The means at visit #3 (78.58 ± 7.51 mmHg) and visit #4 (78.02 ± 6.44 mmHg) were significantly higher as compared to visit #5 (74.64 ± 5.30 mmHg). In other words, there was a statistically significant reduction in mean diastolic BP at visit #5 as compared to previous visits. Pulse and heart rate were insignificantly different across visits. The mean respiratory rate showed a significant difference across visits (p- 5 value: 0.033). The mean at visit #3 (18.84 ± 1.96/min) and visit #4 (18.51 ± 1.93/min) were significantly higher than the baseline (18.04 ± 1.51/min). A marginal lowering of mean was observed at visit #5 (18.47 ± 1.90/min) compared to visit #4. The mean temperature was insignificantly different across visits.

Normal physical parameters: The number of patients with normal physical 10 parameters across visits in two study groups was evaluated as mentioned below in table 6:

Table 6:

Inference: It is evident from table 6 that in both groups, all the clinically evaluated 15 patients at each visit had normal physical parameters.

Comparison of anthropometric parameters between two groups (formulation- 252A and placebo-252B) at each visit as per the treatment protocol analysis is depicted in table 7A: Table 7A:

wherein * P-value calculated by using t-test for independent samples and bold p- value indicate statistical significance. Inference: It is evident from table 7A and figure 9A that at baseline, out of 11 parameters, weight (p-value: 0.021), basal metabolic rate (p-value: 0.008), total body water (p-value: 0.03) and lean body mass (p-value: 0.019) showed a statistically

5 significant difference between two groups. The mean values of these parameters were significantly lower in 252 A arm as compared to 252B. The remaining 7 parameters were insignificantly different in the two arms. At visit #3, in addition to the above four parameters, calf circumference (p-value: 0.017) also showed a significantly higher mean in 252B arm as compared to 252A. Further at visit #4,

10 waist circumference (p-value: 0.036), upper mid-arm circumference (p-value: 0.026), calf circumference (p-value: 0.005) indicated a statistically significant difference between the two treatment arms. The other four parameters i.e. Weight, BMR, TBW, and LBM continued showing significant differences between the two arms. At visit #5, the mean hip circumference in arm 252B was significantly higher

15 than the mean of arms 252A (p-value: 0.012). Other parameters like weight (p-value: 0.001), waist circumference (p-value: 0.02), upper mid- arm circumference (p-value: 0.021), calf circumference (p-value: 0.001), BMR (p-value: 0.001), TBW (p-value: 0.005) and LBM (p-value: 0.005) continued showing significant difference between two groups.

20 Comparison of anthropometric parameters between two groups (formulation- 252A and placebo-252B) at each visit as per the protocol analysis is given in table 7B:

The analysis was performed on subjects with complete information till the last follow-up in both the treatment arms.

25 Table 7B:

wherein *P-value calculated by using repeated measure analysis of variance (ANOVA). Bold p-values indicate the statistical significance.

Inference: It is evident from table 7B and figure 9B that at baseline, except basal 5 metabolic rate (p-value: 0.017) and lean body mass (p-value: 0.031), all the other parameters showed an insignificantly different means between two treatment arms. At visit #3, weight (p-value: 0.014), upper mid-arm circumference (p-value: 0.03), calf circumference (p-value: 0.025), BMR (p-value: 0.009), TBW (p-value: 0.031) and LBM (p-value: 0.018) showed significantly different means between two ar s. The means for these parameters were lower in 252 A arm as compared to 252B. Further, at visit #4, waist circumference also showed significant difference between 5 two groups (p-value: 0.036), in addition to weight (p-value: 0.002), upper mid-arm circumference (p-value: 0.026), calf circumference (p-value: 0.005), BMR (p-value: 0.003), TBW (0.014) and LBM (p-value: 0.01). Again, the mean values were lower in arm 252A as compared to 252B. At visit #5, hip circumference also indicated statistically significant difference between two arms (p-value: 0.012). The other 10 parameters like weight (p-value: 0.001), waist circumference (p-value: 0.02), upper mid-arm circumference (p-value: 0.021), calf circumference (p-value: 0.001), BMR (p-value: 0.001), TBM (p-value: 0.005) and LBM (p-value: 0.005) continued showing significant difference between two arms.

Comparison of anthropometric parameters across visits in two treatment 15 groups (formulation-252A and placebo-252B) as per protocol analysis is mentioned in table 8:

Table 8:

Inference: It is evident from table 8 and figure 10 that in arm 252A, all the 5 parameters showed a statistically significant reduction in the mean parametric values at visit #5 as compared to baseline. The mean weight reduced significantly from baseline (78.65 ± 12.13 kg) to visit #5 (74.52 ± 11.71 kg) with a p-value < 0.0001. The mean waist circumference reduced significantly from baseline (98.85 ± 10.09 cm) to visit #5 (95.11 ± 9.16 cm) with a p-value < 0.0001. Further, hip 10 circumference reduced significantly from 108.93 ± 9.71 cm at baseline to 102.59 ± 10.35 cm at visit #5 with a p-value < 0.0001. The waist-hip ratio showed statistically significant increase from baseline (0.91 ± 0.1) to visit #5 (0.93 ± 0.11) with associated p-value of 0.021. The mean neck circumference showed significant lowering from baseline (36.61 + 3.5 cm) to visit #5 (35.61 + 3.10 cm) with a p-value < 0.0001. Similarly, upper mid- arm circumference reduced from 32.3 ± 3.15 cm at baseline to 31.39 ± 3.22 cm at visit #5 with a p-value < 0.0001. The mean calf circumference also reduced from 38.8 ± 0.3 cm to 36.54 ± 3.12 cm with a p-value < 0.0001. The body fat % also showed significant drop from 40.72 ± 10.4% to 37.39 ±

9.43% with a p-value < 0.0001. The BMR at baseline (1476.69 ± 247.68 cal/day) showed significant reduction at visit #5 (1436.13 ± 240.31 cal/day) with a p-value < 0.0001. Further, total body water showed a significant reduction to 36.04 ± 7.11% at visit #5 from 37.28 ± 7.56% at baseline with a p-value < 0.0001. The mean lean body mass also reduced significantly from 49.44 ± 10.15 kg at baseline to 48.22 ± 9.57 kg at visit #5 with a p-value < 0.0001.

In arm 252B, only three parameters viz., hip circumference, neck circumference, and calf circumference showed a statistically significant reduction of the mean from baseline to visit #5, each with a p-value < 0.0001. Comparison of the percent reduction in anthropometric parameters from baseline to visit #5 between two treatment arms (formulation arms-252A) is mentioned in table 8A

Table 8A:

wherein *P- value is calculated using t-test for independent samples and bold p-value indicate statistical significance.

Inference: It is evident from Table 8 A that the mean reduction in weight, waist and 5 hip circumference was significantly higher in the formulation group 252A as compared to the placebo group 252B (p-value < 0.0001). Further, neck circumference was also significantly reduced in the formulation group as compared to the placebo group (p-value =0.013). Other anthropometric parameters like calf- circumference, body fat, BMR, TBW, and LBM also showed significantly higher 10 reduction in the polyherbal formulation group as compared to the placebo group (p- value < 0.0001).

Comparison of hematological parameters between two groups (formulation- 252A and placebo- 252B) at screening and visit #5 is mentioned in table 9

Table 9:

wherein * P-value is calculated by using t-test for independent samples

Inference: It is evident from table 9 that at screening, none of the parameters showed a statistically significant difference between the two treatment arms (p-value 5 > 0.05). Similarly, the analysis at visit #5 indicated a statistically insignificant difference of hematological parameters between the two treatment arms (p-value > 0.05).

Comparison of lipid parameters between two groups (formulation-252A and 10 placebo-252B) at different visits is mentioned in table 10

Table 10:

value indicate statistical significance.

Inference: It is evident from table 10 and figure 11 that at baseline and visit #3, all 5 the parameters showed a statistically insignificant difference of means between two arms (p-value > 0.05). Further, at visit #4, only mean triglycerides in arm 252A (205.16 ± 86.81 mg/dl) were significantly higher than that of arm 252B (164.22 ± 67.34 mg/dl) with a p-value of 0.007.

Comparison of lipid parameters across visits in two treatment groups 10 (formulation-252A and placebo-252B) as mentioned in table 11

Table 11:

wherein *P-value is calculated by using repeated measure analysis of variance (ANOVA). Bold p-values indicate the statistical significance.

Inference: It is evident from table 11 and figure 12 that in treatment arm 252 A, the mean HDL showed a statistically significant increase from baseline (39.57 ± 7.04 mg/dl) to visit #5 (46.30 ± 20.5 mg/dl) as indicated by the p-value of 0.009. Further, LDL also showed statistically significant reduction from baseline (127.47 ± 32.38 mg/dl) to visit #5 (111.94 ± 32.33 mg/dl) with a p-value of 0.012. However, the mean VLDL increased from 16.44 ± 8.49 mg/dl at baseline to 21.64 ± 12.45 mg/dl at visit #5 with a p-value of 0.014. In arm 252B, LDL showed a statistically significant reduction from 122.78 ± 30.25 mg/dl at baseline to 114.18 ± 26.05 mg/dl at visit #5 with a p-value of 0.037. Other parameters showed an insignificant change of mean values across visits.

Conclusion:

• A total of 130 patients were screened for the trial out of which 120 patients were randomized to two treatment groups equally viz. the active group (252A: formulation of the present disclosure) and the placebo group (252B). At visit #3, 113 patients were evaluated, while 7 patients were lost to follow up. Further, at visit #4, 4 patients were lost and thus finally 109 patients were evaluated at visit #5.

• The vital parameters differed insignificantly between two groups in treatment analysis at baseline, visit #3, visit #4 and visit #5, except diastolic BP, which was significantly lower in group placebo group-252B as compared to the formulation of the present disclosure group-252A (p-value: 0.025). Similar results were observed as per-protocol analysis.

• Within-group analysis for vital parameters revealed that the formulation of the present disclosure -252A showed statistically significant change across visits which is not clinically significant with respect to pulse (p-value: 0.003) and respiratory rate (p-value: 0.001). In placebo group-252B, diastolic BP (p- value: 0.003) showed statistically significant change across visits, which again was clinically insignificant. The respiratory rate also showed significant change across time (p-value: 0.033).

• At visit #5, the anthropometric parameters like weight (p-value: 0.001), waist circumference, hip circumference (p-value: 0.02), upper mid-arm circumference (p-value: 0.021), calf- circumference (p-value: 0.001), basal metabolic rate (p-value: 0.001), total body water (p- value: 0.005) and lean body mass (p-value: 0.005) had significantly lower mean in the formulation group-252 A as compared to placebo group-252B.

• The change in anthropometric measurements across visits was studied in each treatment arm. In formulation group- 252A, all the parameters showed statistically significant change (reduction) in the mean values across visits (p- value < 0.0001). However, in the placebo group, only hip, neck, and calf circumferences showed statistically significant differences across visits (p- value < 0.0001).

• Also, the percent reduction of these parameters from baseline till visit #5 was obtained and compared between two treatment arms. The mean reduction in weight, waist and hip circumference was significantly higher in the formulation group - 252A as compared to the placebo group- 252B (p-value < 0.0001). Further, neck circumference was also significantly reduced in the formulation group as compared to the placebo group (p-value =0.013). Other anthropometric parameters like calf-circumference, body fat, BMR, TBW, and LBM also showed significantly higher reduction in the formulation group as compared to the placebo group (p-value < 0.0001).

• The comparison of each lipid parameter was performed between two treatment arms at each visit. All the parameters indicated a statistically insignificant difference of means between two arms in each visit.

• The comparison of each lipid parameter was also performed in each treatment arm across visits. In the formulation group, parameters like HDL (p-value: 0.009), LDL (p-value: 0.012), and VLDL (p-value: 0.014) showed statistically significant change in the mean values from baseline to visit #5, while in the placebo group, only LDL showed significant change in the mean values from baseline to visit #5 (p- value: 0.037).

EXPERIMENT 4: Comparative examples:

Table 12:

5 Inference: It is evident from table 12 that the formulation of the present disclosure is present in very low amounts in comparison to the conventional formulations as reported in the prior art. Surprisingly, the contents of active ingredients in the present formulation are several folds less than what is reportedly used by conventional formulations indicating that the present formulation has improved properties of anti-obesity as compared to individual extracts reported in the literature.

TECHNICAL ADVANCES AND ECONOMICAL SIGNIFICANCE

The polyherbal formulation of the present disclosure described herein above has several technical advantages including, but not limited to, the realization of; a polyherbal formulation has a better anti-obesity and anti-diabetic properties.

The embodiments as described herein above, and various features and advantageous details thereof are explained with reference to the non-limiting embodiments in the following description.

Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

The use of the expression “at least” or “at least one” suggests the use of one or more elements or ingredients or quantities, as the use may be in the embodiment of the disclosure to achieve one or more of the desired objects or results.

The foregoing description of specific embodiments so fully reveal the general nature of the embodiments herein, that others can, by applying current knowledge, readily modify and/or adapt for various applications of such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the embodiments as described herein. Further, it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.

Having described and illustrated the principles of the present disclosure with reference to the described embodiments, it will be recognized that the described embodiments can be modified in arrangement and detail without departing from the scope of such principles.

While considerable emphasis has been placed herein on the components and component parts of the preferred embodiments, it will be appreciated that many embodiments can be made and that many changes can be made in the preferred embodiments without departing from the principles of the disclosure. These and other changes in the preferred embodiment as well as other embodiments of the disclosure will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.

Claims

CLAIMS:

1. A polyherbal formulation comprising;

(a) an extract of Coffee arabica in an amount in the range of 15 wt.% to 20 wt.% with respect to the total weight of the formulation; wherein said extract of coffee arabica has at least 13% of chlorogenic acid;

(b) an extract of Garcinia cambogia in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation; wherein said extract of Garcinia cambogia has at least 50% of hydroxycitric acid;

(c) an extract of Terminalia arjuna in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation; wherein said extract of Terminalia arjuna has at least 50% of polyphenols;

(d) an extract of Commiphora mukul in an amount in the range of 2 wt.% to 10 wt.% with respect to the total weight of the formulation; wherein said extract of Commiphora mukul has at least 0.5% of guggulsterone E and guggulsterone Z;

(e) an extract of Cyperus rotundus in an amount in the range of 5 wt.% to 15 wt.% with respect to the total weight of the formulation; wherein said extract of Cyperus rotundus has at least 5% of polyphenols;

(f) an extract of Zingiber officinale in an amount in the range of 0.5 wt.% to 5 wt. % with respect to the total weight of the formulation; wherein said extract of Zingiber officinale has at least 0.4% of polyphenols;

(g) an extract of Piper longum in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation; wherein said extract of Piper longum has at least 1% of piperine;

(h) an extract of Piper nigrum in an amount in the range of 0.5 wt.% to 5 wt.% with respect to the total weight of the formulation; wherein said extract of Piper nigrum has at least 2% of piperine; and (i) a pharmaceutically acceptable excipient.

2. The polyherbal formulation as claimed in claim 1, wherein said pharmaceutically acceptable excipient is selected from the group consisting of a disintegrant, a binder, a preservative, a fluid medium, a glidant, a film-forming agent, and a coloring agent.

3. The polyherbal formulation as claimed in claim 2, wherein said disintegrant is at least one selected from the group consisting of polyvinylpyrrolidone (crospovidone), sodium starch glycolate, croscarmellose sodium, starch, and colloidal anhydrous silica; wherein the amount of said disintegrant is in the range of 15 to 20% with respect to the total weight of the formulation.

4. The polyherbal formulation as claimed in claim 2, wherein said binder is at least one selected from the group consisting of microcrystalline cellulose, povidone, hydroxymethyl cellulose, ethyl cellulose, hydroxypropyl cellulose, starch, gum acacia, alginate, and carboxymethyl cellulose; wherein the amount of said binder is in the range of 20 to 25% with respect to the total weight of the formulation.

5. The polyherbal formulation as claimed in claim 2, wherein said preservative is at least one selected from the group consisting of methyl hydroxybenzoate (methylparaben), propyl hydroxybenzoate (propylparaben), ethyl paraben, and sodium benzoate; wherein the amount of said preservative is in the range of 0.01 to 0.05% with respect to the total weight of the formulation.

6. The polyherbal formulation as claimed in claim 2, wherein said fluid medium is at least one selected from the group consisting of isopropanol (isopropyl alcohol), polyethylene glycol 4000, and dichloromethane (methylene chloride).

7. The polyherbal formulation as claimed in claim 2, wherein said glidant is at least one selected from the group consisting of silicon dioxide a, magnesium stearate, talc, calcium palmitate, ascorbyl palmitate, and starch; wherein the amount of said glidants is in the range of 1 to 5% with respect to the total weight of the formulation.

8. The polyherbal formulation as claimed in claim 2, wherein said film-forming agent is at least one selected from the group consisting of hydroxypropyl methylcellulose, ethyl cellulose, hydroxypropyl cellulose, povidone, and polyvinyl alcohol; wherein the amount of said film-forming agent is in the range of 1 to 5% with respect to the total weight of the formulation.

9. The polyherbal formulation as claimed in claim 2, wherein said coloring agent is at least one selected from the group consisting of titanium dioxide, iron oxide red, iron oxide black, tartrazine yellow, indigotine, sunset yellow, and brilliant blue; wherein the amount of said coloring is in the range of 0.1 to 0.5% with respect to the total weight of the formulation.

10. The polyherbal formulation as claimed in claim 1, said formulation comprises: a) an extract of Coffee arabica in an amount in the range of 17 wt.% to 19 wt.% with respect to the total weight of the formulation; wherein said extract of coffee Arabica has at least 13% of chlorogenic acid; b) an extract of Garcinia cambogia in an amount in the range of 7 wt.% to 12 wt.% with respect to the total weight of the formulation; wherein said extract of Garcinia cambogia has at least 50% of hydroxycitric acid; c) an extract of Terminalia arjuna in an amount in the range of 6 wt.% to 10 wt.% with respect to the total weight of the formulation; wherein said extract of Terminalia arjuna has at least 50% of polyphenols; d) an extract of Commiphora mukul in an amount in the range of 4 wt.% to 8 wt. % with respect to the total weight of the formulation; wherein said extract of Commiphora mukul has at least 0.5% of guggulsterone E and Z; e) an extract of Cyperus rotundus in an amount in the range of 6 wt.% to 10 wt.% with respect to the total weight of the formulation; wherein said extract of Cyperus rotundus has at least 5% of polyphenols; f) an extract of Zingiber officinale in an amount in the range of 1 wt.% to 3 wt.% with respect to the total weight of the formulation; wherein said extract of Zingiber officinale has at least 0.4% of polyphenols; g) an extract of Piper longum in an amount in the range of 1 wt.% to 3 wt.% with respect to the total weight of the formulation; wherein said extract of Piper longum has at least 1% of piperine; h) an extract of Piper nigrum in an amount in the range of 1 wt.% to 3 wt.% with respect to the total weight of the formulation; wherein said extract of Piper nigrum has at least 2% of piperine; and i) a pharmaceutically acceptable excipient.

11. The polyherbal formulation as claimed in claim 1, wherein said formulation is in a form selected from tablet, uncoated tablet, film-coated tablet, sugar-coated tablet, and enteric-coated tablet.

12. The polyherbal formulation as claimed in claim 11, wherein said tablet is film- coated tablet.

13. A process for the preparation of the polyherbal formulation wherein said polyherbal formulation is in the form of a tablet, said process comprising the following steps: a) sifting individually and then mixing, predetermined amounts of sieved powder extracts of coffee arabica, garcinia cambogia, terminalia arjuna, commiphora mukul, cyperus rotundus, zingiber officinale, piper longum and piper nigrum with predetermined amounts of a binder, a glidant, and a disintegrant in a mixer under stirring to obtain a first mixture; b) separately, mixing a predetermined amount of at least one fluid medium and a predetermined amount of at least one preservative in a vessel to obtain a solution; c) adding said solution of step (b) into said first mixture obtained in step (a) in the mixer under stirring for a first predetermined time period to obtain a second mixture; d) kneading said second mixture under stirring for a second predetermined time period followed by granulation to obtain granules; e) drying said granules at a predetermined temperature to obtain a dried granules; f) milling said dried granules to obtain a milled granules; g) mixing said milled granules with a predetermined amount of at least one disintegrant and a predetermined amount of at least one binder for a third predetermined time period to obtain a blend; h) mixing the blend with a predetermined amount of at least one glidant for a fourth predetermined time period to obtain a lubricated granules; and i) punching the lubricated granules to obtain the polyherbal formulation in the form of tablet.

14. The process as claimed in claim 13, wherein said predetermined amount of extract of Coffee arabica is in the range of 15 wt.% to 20 wt.%, said predetermined amount of extract of Garcinia cambogia is in the range of 5 wt.% to 15 wt.%, said predetermined amount of extract of Terminalia arjuna is in the range of 5 wt.% to 15 wt.%, said predetermined amount of extract of Commiphora mukul is in the range of 2 wt.% to 10 wt.%, said predetermined amount of extract of Cyperus rotundas is in the range of 5 wt.% to 15 wt.%, said predetermined amount of extract of Zingiber officinale is in the range of 0.5 wt.% to 5 wt.%, said predetermined amount of extract of Piper longum is in the range of 0.5 wt.% to 5 wt.%, said predetermined amount of extract of Piper nigrum is in the range of 0.5 wt.% to 5 wt.%, said predetermined amount of binder is in the range of 20 to 25% with respect to the weight of the formulation, said predetermined amount of glidant is in the range of 1 to 5% with respect to the weight of the formulation, said predetermined amount of disintegrant is in the range of 15 to 20% with respect to the weight of the formulation, and said predetermined amount of preservative is in the range of 0.01 to 0.05% with respect to the weight of the formulation.

15. The process as claimed in claim 13, wherein said tablet is coated with a layer of a predetermined amount of at least one coating solution, followed by adding a predetermined amount of at least one glidant and a coloring agent to obtain a film-coated tablet.

16. The process as claimed in claim 13, wherein said first predetermined time period is in the range of 3 to 6 min, said second predetermined time period is in the range of 4 to 8 min, said third predetermined time period is in the range of 8 to 12 min and said fourth predetermined time period is in the range of 2 to 4 min.

17. The process as claimed in claim 13, wherein said predetermined temperature is in the range of 40 to 50°C.

18. The process as claimed in claim 15, wherein said coating solution comprises hydroxypropyl methylcellulose, polyethylene glycol, titanium dioxide, talc, colors, and solvents.